畜牧与饲料科学 ›› 2010, Vol. 31 ›› Issue (9): 4-4.doi: 10.12160/j.issn.1672-5190.2010.09.003

• 基础科学 • 上一篇    下一篇

猪群中7种高发传染病多重PCR诊断方法的建立

贺显晶 孙东波 周玉龙 林云成 韩旭 王越强 郭婷婷 武瑞   

  1. 黑龙江八一农垦大学动物科技学院,黑龙江大庆163319
  • 出版日期:2010-09-20 发布日期:2010-09-20
  • 通讯作者: 贺显晶
  • 作者简介:贺显晶(1985-),女,助教,硕士,主要研究方向为动物疫病诊断。 通讯作者:孙东波(1978-),男,博士,主要研究方向为动物疾病诊断与药物开发。
  • 基金资助:
    黑龙江省普通高等学校青年学术骨干支持计划项目(1154G60).

Development of Multiplex PCR Method for Detection of Seven Common Infectious Diseases in Pigs

HE Xian-jing, SUN Dong-bo, ZHOU Yu-long, LIN Yun-cheng, HAN Xu, WANG Yue-qiang, GUO Ting-ting, WU Rui (College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing 163319, China)   

  • Online:2010-09-20 Published:2010-09-20

摘要: [目的]建立猪瘟、蓝耳病(又称猪繁殖与呼吸综合征)、圆环病毒2型感染、猪链球菌病、副猪嗜血杆菌病、猪多杀性巴氏杆菌感染和猪支原体感染7种猪病的多重PCR诊断方法。[方法]根据GenBank发表的核苷酸序列设计了分别针对猪瘟、蓝耳病、圆环病毒2型感染、猪链球菌病、副猪嗜血杆菌病、猪多杀性巴氏杆菌感染和猪支原体感染的2套多重PCR特异性引物,优化多重PCR反应条件后进行敏感性与特异性评价。[结果]建立的2套多重PCR诊断方法对其他常见猪病病原的基因组无特异性扩增,检测病原基因组最低浓度可达到pg级。[结论]该研究建立的2套多重PCR诊断方法可以用于猪群中上述7种猪病痛原的单一感染或混合感染的鉴别诊断。

Abstract: [Objective] To develop multiplex PCR method for detection of classical swine fever, porcine reproductive and respiratory syndrome, porcine circovirus type 2, Streptococcus suis, Haemophilus parasuis, swine Pasteurella multocida and swine Mycoplasma hyopneumonia. [Method] Seven pairs of specif/c primers were designed for two multiplex PCR based on the conservative nucleotide sequences published in GenBank. After optimizing reaction conditions, sensitivity and specificity of two multiplex PCR methods were evaluated, respectively. [Result] The genomic DNA of other common pathogens were not amplified by the two multiplex PCR methods, and the detection limit of the two multiplex PCR methods was at a picogram level of genomic DNA or cDNA. [Conclusion] The two multiplex PCR methods can be used for diagnosis of single infection or mixed infection of above seven pathogens.

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